Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free
See all Apolipoprotein E4 primary antibodies
Description

Rabbit monoclonal [EPR24181-64] to Apolipoprotein E4 - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK-293T transfected with APOE4 (C130R) expression vector containing a myc-His-tag®; Human plasma. IHC-P: HEK-293T transfected with APOE4 (C130R); Human placenta (plasma) , breast cancer and kidney tissue. ICC: 293T. Flow cyt: 293T transfected with myc-tagged APOE4. IP: HEK-293T transfected with myc-tagged APOE4.
General notes
ab279719 is the carrier-free version of ab279714. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab279719 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C.
Storage buffer

Constituent: 100% PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR24181-64
Isotype

IgG
Research areas

Western blot - Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free (ab279719)

Anti-Apolipoprotein E4 antibody [EPR24181-64] (ab279714) at 1/1000 dilution + Human plasma at 5 µl

Secondary
Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 36 kDa
Observed band size: 36 kDa

This data was developed using ab279714, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Exposure time: 3 minutes
Western blot - Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free (ab279719)

All lanes : Anti-Apolipoprotein E4 antibody [EPR24181-64] (ab279714) at 1/1000 dilution

Lane 1 : HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293T transfected with Apolipoprotein E2 (R176C) expression vector containing a myc-His-tag®, whole cell lysate
Lane 3 : HEK-293T transfected with Apolipoprotein E3 expression vector containing a myc-His-tag®, whole cell lysate
Lane 4 : HEK-293T transfected with Apolipoprotein E4 (C130R) expression vector containing a myc-His-tag®, whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 36 kDa
Observed band size: 40 kDa
why is the actual band size different from the predicted?

This data was developed using ab279714, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Exposure time: 3 minutes
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free (ab279719)

This data was developed using ab279714, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Apolipoprotein E4 with ab279714 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human kidney. (PMID:29692990 ) The section was incubated with ab279714 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free (ab279719)

This data was developed using ab279714, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Apolipoprotein E4 with ab279714 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human breast cancer. The section was incubated with ab279714 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free (ab279719)

This data was developed using ab279714, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human placenta (plasma) tissue labeling Apolipoprotein E4 with ab279714 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the plasma in human placenta. (PMID: 26828652) The section was incubated with ab279714 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free (ab279719)

This data was developed using ab279714, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded sections Panel A and D: HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with APOE4 (C130R) expression vector containing a myc-His-tag®
Panel B: HEK-293T transfected with APOE2 (R176C) expression vector containing a myc-His-tag®. Panel C: HEK-293T transfected with APOE3 expression vector containing a myc-His-tag® labeling Apolipoprotein E4 with ab279714 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) HEK-293T transfected with an APOE4 (C130R) expression vector is observed. Almost no staining on (B) HEK-293T transfected with an APOE2 (R176C) expression vector and (C) HEK-293T transfected with APOE3 expression vector. (D) Secondary antibody only control for (A). The section was incubated with ab279714 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunocytochemistry - Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free (ab279719)

This data was developed using ab279714, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling Apolipoprotein E4 with ab279714 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing strong cytoplasmic staining in 293T cells transfected with APOE4-Myc plasmid is observed. Myc-Tag Mouse mAb (Alexa Fluor® 647) antibody was used to counterstain transfected proteins at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Immunocytochemistry - Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free (ab279719)

This data was developed using ab279714, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling Apolipoprotein E4 with ab279714 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution. Negative control: Confocal image showing no staining in 293T cells transfected with APOE2-Myc or APOE3-Myc plasmid.. Myc-Tag Mouse mAb (Alexa Fluor® 647) antibody was used to counterstain transfected proteins at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Flow Cytometry - Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free (ab279719)

This data was developed using ab279714, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell) transfected with myc-tagged Apolipoprotein E4 construct cells labeling Apolipoprotein E4 with ab279714 at 1/500 dilution (Right panel) compared with Rabbit monoclonal IgG Isotype Control (ab172730) (Left panel). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry - Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free (ab279719)

This data was developed using ab279714, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell) transfected with myc-tagged Apolipoprotein E2 construct (Left)/ 293T transfected with myc tagged Apolipoprotein E3 construct (Right) labeling Apolipoprotein E4 with ab279714 at 1/500 dilution. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-Apolipoprotein E4 antibody [EPR24181-64] - BSA and Azide free (ab279719)

This data was developed using ab279714, the same antibody clone in a different buffer formulation.

Apolipoprotein E4 was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney) transfected with Apolipoprotein E4 (C130R) expression vector containing a myc-His-tag® whole cell lysate 10 μg with ab279714 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab279714 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1:HEK-293T (human embryonic kidney) transfected with Apolipoprotein E4 (C130R) expression vector containing a myc-His-tag® whole cell lysate 10 μg

Lane 2: ab279746 IP in HEK-293T (human embryonic kidney) transfected with Apolipoprotein E4 (C130R) expression vector containing a myc-His-tag® whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab279714 in HEK-293T transfected with Apolipoprotein E4 (C130R) expression vector containing a myc-His-tag® whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 41 seconds